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Mycoplasma PCR Detection Kit

 

CAT.NO UNIT
G238 100rxn
Description The Mycoplasma PCR Detection Kit offers high specificity and sensitivity to minimize false positives while ensuring coverage over 200 strains of Mycoplasmas for quick and reliable routine screening of cell cultures less than 2 hours. Mycoplasmas are highly undesirable, easily acquired and notorious for an elusive onset of infection. It can alter the infected cells at a molecular level leading to visible changes in cell morphology and growth characteristics. Timely detection of Mycoplasmas in cell cultures is recommended in order to deter wide-spread contamination and save on the costly efforts of elimination. The MasterMix contains gel loading dye, making it convenient to load for gel electrophoresis.

 

PRODUCT COMPONENT QUANTITY
BlasTaq™ 2X PCR MasterMix
1.25 ml
Primer Mix
100 μl
Positive Control
250 μl
Nuclease-Free H2O
1.0 ml
SKU G238
Storage Condition

Store at -20°C.
Unit quantity 100 rxn

Supporting Protocol

MSDS

QC
Other

Yes, we do have a positive control included in the kit.

We recommend culturing your cells in a 6-well plate or 10cm dish.

It is DNA fragments of Mycoplasma gDNA diluted in medium.

yes

It is not necessary to remove routine antibiotics (e.g. penicillin, streptomycin, and gentamicin) from the media prior to PCR detection, as they will not interfere with the assay.

Our kit is able to detect as low as 10 copies of Mycoplasma /sample

Mycoplasma testing is more dependent on how recently the cells have been subcultured. Seeding density does not play a major role when using this kit. We recommend cells be around at least 3-5 days old in culture without changing any fresh media so as to detect secreted mycoplasma in culture supernatant (i.e. the cell culture medium should not be refreshed/changed for 48-72 hours prior to collection).

Using samples stored at -80C should be fine for this kit. If you thawing out the cells and performing mycoplasma testing right away, usually when thawing the cells the supernatant from the vial is diluted when adding to the cell culture vessel. As long as the DMSO is diluted and the DMSO content is less than 0.5% final in the PCR reaction it should not cause a problem. Too much DMSO would alter the Tm of the PCR buffer and thus affect amplification.

In general, the more fresh the sample is when running the PCR, the better; in keeping with this, it is normally advised to wait until all cell lines are ready to go prior to collecting the supernatant for each. However, if necessary, you can certainly store collected supernatant at +4C storage for short-term (1-2 days) storage; note that we highly recommend avoiding repeated freeze thaw cycles.

We recommend the cells should remain in culture for at least 48-72 hours prior to screening for the presence of mycoplasmas, and that the media sample collection only be done once the cells have reached at least 80% confluence.

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